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. 2021 Nov 26;24(12):103517. doi: 10.1016/j.isci.2021.103517

Figure 1.

Figure 1

GFAT2 is a major GFAT isoform in the heart and cardiomyocytes

(A) mRNA expression levels of GFAT1 and GFAT2 adjusted by18S mRNA in lung, liver, brain, heart, testis, and kidney (n = 3).

(B) Representative immunoblots of GFAT1, GFAT2, and β-actin and Coomassie brilliant blue (CBB) staining in lung, liver, brain, heart, testis, and kidney. Quantitative analysis of the absolute value of intensity of GFAT1 and GFAT2 in lung, liver, brain, heart, testis, and kidney (n = 3).

(C) mRNA expression levels of GFAT1 and GFAT2 adjusted by18S mRNA in ISO (15 mg/kg body weight/day, 7 days)- or control vehicle (saline)-treated C57B/6J mouse hearts (n = 5).

(D) Representative immunoblots of GFAT1, GFAT2, OGT, OGA, protein O-GlcNAcylation, and GAPDH in ISO (15 mg/kg body weight/day, 7 days)- or control vehicle (saline)-treated C57B/6J mouse hearts. Quantitative analysis of GFAT1, GFAT2, OGT, OGA, and protein O-GlcNAcylation in ISO- or control vehicle-treated C57B/6J mouse hearts (n = 5).

(E) Representative immunoblots of GFAT1, GFAT2, and β-actin and CBB staining in neonatal rat ventricular cardiomyocytes (NRVMs), fibroblasts (FBs), endothelial cells (ECs), C2C12 cells, and HEK293 cells. Quantitative analysis of the absolute value of intensity of GFAT1, GFAT2 in NRVMs, FBs, ECs, C2C12 cells, and HEK293 cells (n = 3).

(F) Representative immunoblots of GFAT1, GFAT2, OGT, OGA, protein O-GlcNAcylation, and GAPDH in NRVMs treated with ISO (1 μM) at the indicated time point (n = 6). Quantitative analysis of GFAT1, GFAT2, OGT, OGA, and protein O-GlcNAcylation in NRVMs treated with ISO (1 μM) at the indicated time point (n = 6). ∗p < 0.05, ∗∗p < 0.01: post hoc Tukey's comparison test.